Difference between revisions of "Team:AHUT China/InterLab"

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                 <div align="center"><h4 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;1 . HP--Creative Art Course for primary school students</h4></div>
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                 <div align="center"> <h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Introduction</h2></div>
                   <p>AHUT_China(This year's project is about carbon dioxide.)With the increasing impact of the greenhouse effect on the global environment, it’s very important to know how to efficiently deal with greenhouse gas CO2. Traditional CO2 collection technology is not mature, restricted by high energy consumption and low efficiency. Our project simulates the protein molecule expressed by the mutation of carbonic anhydrase gene Loci, obtains the high thermal stability of carbonic anhydrase (CA-2) and imports it into Escherichia coli cells to achieve efficient CO2 collection</p><br>
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                   <p>We took part in the Fifth International InterLab Measurement Study which ains to achieve the purpose of comparative measurement. The goal of this study is to obtain large amounts of data from labs across the world,to develop absolute units for measurements GFP in a plate reader to eliminate variation between labs.</p><br>
<div align="center"><img src="https://static.igem.org/mediawiki/2018/a/a9/T--AHUT_China--The_Xiangxiangle_art_%26_culture_House.jpg" width="452" height="266"></div><br>
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<div align="center"><h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Materials</h2></div>
                <div align="center"><span style="color: rgba(2,2,2,1)">Figure 1  The Xiangxiangle art & culture House</span></div>
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                  <p>Plate reader: Synergy H1 (Biotek)<br>
  <p><br>AHUT_China(This year's project is about carbon dioxide.)With the increasing impact of the greenhouse effect on the global environment, it’s very important to know how to efficiently deal with greenhouse gas CO2. Traditional CO2 collection technology is not mature, restricted by high energy consumption and low efficiency. Our project simulates the protein molecule expressed by the mutation of carbonic anhydrase gene Loci, obtains the high thermal stability of carbonic anhydrase (CA-2) and imports it into Escherichia coli cells to achieve efficient CO2 collection<br>
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Plate reader plates: Corning 3603 96-Well Microplates (black plates with clear flat bottom)<br>
<div align="center"><img src="https://static.igem.org/mediawiki/2018/a/a2/T--AHUT_China--learn_knowledge.jpg" width="452" height="266" alt=""/></div> <br>
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Cell culture shaker: ZWYR-200D<br><br>
<div align="center">Figure 2  Learning biological knowledge</div>
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Devices:<br>
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Negative control :BBa_R0040 <br>
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Positive control :BBa_I20270 <br>
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Device 1: BBa_J364000  <br>
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Device 2: BBa_J364001  <br>
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Device 3: BBa_J364002  <br>
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Device 4: BBa_J364007  <br>
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Device 5: BBa_J364008  <br>
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Device 6: BBa_J364009  <br>
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Note: for Device 5, we have not transformed it into DH5⍺ competent cells successfully for many times, therefore, we thank IGEM team of Nanjing University for providing the Device 5.<br>
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Calibration material: Provided in the 2018 IGEM distribution kit <br>
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Microorganism: Escherichia coli DH5⍺ strains<br>
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<div align="center"><h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Methods</h2></div>
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                  <p>Following iGEM requirements, Team AHUT_China performed measurements according to these 2018 InterLab Protocols https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf </p><br>
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<div align="center"><h2 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results</h2></div>
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                  <p>1. OD 600 reference point<br>
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Using OD 600 and H2O to generate the conversion factor for the transformation later. The average of OD600 is 0.063; the correction factor (OD600/ABS600) is 3.500
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  <div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/LUDOX correct result_meitu_11.jpg" width="317" height="234" alt=""/></div><br><div align="center">Fig. 1 LUDOX correct value
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                  <p>2.Particle standard curve<br>
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We obtained the two Particle Standard Curve (normal and log scale).
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  <div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 2 Particle Standard Curve_meitu_2.jpg" width="701" height="440" alt=""/></div><br><div align="center">Fig. 2 Particle Standard Curve
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<p>2.Particle standard curve<br>
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We obtained the two Particle Standard Curve (normal and log scale).
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<div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 3 Particle Standard Curve (log scale)_meitu_3.jpg" width="701" height="440" alt=""/></div><br><div align="center">
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  Fig. 3 Particle Standard Curve (log scale)
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<p>3.Fluorescein standard curve<br>
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Dilution serious of fluorescein were prepared and measured in a 96 well plate. A standard curve is generated to correct the cell based readings to an equivalent fluorescein concentration.<br>
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We obtained the two Fluorescein Standard Curve (normal and log scale).
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<div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 4 Fluorescein Standard Curve_meitu_4.jpg" width="701" height="440" alt=""/></div><br><div align="center">
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  Fig. 4 Fluorescein Standard Curve
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<div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 5 Fluorescein Standard Curve (log scale)_meitu_5.jpg" width="701" height="440" alt=""/></div><br><div align="center">
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  <p align="center" >Fig. 5 Fluorescein Standard Curve (log scale) </p>
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  <p>4.Cell measurements </p>
<p>On July 5, 2018, we arrived at the XiangXiangle Art and Culture House, and we had a creative art class with the children. In class, we studied biology with them and learned creative art courses.
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By letting children learn the signs of iGEM competition and explaining biological knowledge for them, especially the concepts of genes and synthetic biology, etc., we encouraged children to add their own ideas to the iGEM logo and use painting to depict genes, synthetic biology, and iGEM competitions in their heart. As shown in Picture 3.</p><br> <div align="center"><img src="https://static.igem.org/mediawiki/2018/c/c6/T--AHUT_China--The_process_of_class1.jpg" width="192" height="239" alt=""/>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/f/f2/T--AHUT_China--The_process_of_class2.jpg" width="192" height="239" alt=""/>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/f/f0/T--AHUT_China--The_process_of_class3.jpg" width="192" height="239" alt=""/>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/c/c9/T--AHUT_China--The_process_of_class4.jpg" width="192" height="239" alt=""/></div>
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<p>&nbsp;</p><br>
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<div align="center">Picture 3  The process of class</div><br>
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<div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 6 Fluorescence Measurements Curve _meitu_11.jpg" width="732" height="492" alt=""/></div><br><div align="center">
<p>After learning the course we designed, every child completed their own works with the guidance of the teacher Long Lijuan and the iGEM team members.</p><br>
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  <p >Fig. 6 Fluorescence Measurements Curve</p>
<div align="center"><img src="https://static.igem.org/mediawiki/2018/3/37/T--AHUT_China--_Works_completed1.jpg" width="192" height="239" alt=""/>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/8/8c/T--AHUT_China--_Works_completed2.jpg" width="192" height="239" alt=""/>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/a/af/T--AHUT_China--_Works_completed4.jpg" width="192" height="239" alt=""/>&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/d/da/T--AHUT_China--_Works_completed3.jpg" width="192" height="239" alt=""/></div>
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    <p>Test devices 1 and 4 show high fluorescence intensity. Test devices 2 show a modest fluorescence intensity alone with positive control group, while devices3,5,6 barely show low fluorescence intensity alone with the negative control group.
<div align="center">Picture 4  Works completed<br></div><br>
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/c/ce/T--AHUT_China--_Group_photo1.jpg" width="304" height="234" alt=""/>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/c/ce/T--AHUT_China--_Group_photo2.jpg" width="303" height="232" alt=""/></div>
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<div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 7 Raw OD600 Curve _meitu_7.jpg" width="724" height="484" alt=""/></div><br><div align="center">
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  <p >Fig. 7 Raw OD600 Curve</p>
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<div align="center">Picture 5 Group photo</div><br>
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    <p>5.We obtained the Colony Forming Units per 0.1 OD600 E. coli cultures
  <div align="center"><img src="https://static.igem.org/mediawiki/2018/5/5e/T--AHUT_China--_Single_photo1.jpg" width="173" height="201" alt=""/>&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/e/ef/T--AHUT_China--_Single_photo2.jpg" width="173" height="201" alt=""/>&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/8/82/T--AHUT_China--_Single_photo3.jpg" width="173" height="201" alt=""/>&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/5/50/T--AHUT_China--_Single_photo4.jpg" width="173" height="201" alt=""/>&nbsp;&nbsp;<img src="https://static.igem.org/mediawiki/2018/c/cb/T--AHUT_China--_Single_photo5.jpg" width="173" height="201" alt=""/></div><br>
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<div align="center">Picture 6  Single photo</div><br>
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<p>Our Human Practice-- Creative Art Courses in Primary School Students is very meaningful. It not only introduces the iGEM competition, but also introduces the iGEM team of the Anhui University of Technology. And at the same time, this activity guides the children to have a preliminary understanding of life and the corresponding disciplines.
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<div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 8 CFU Result.png" width="724" height="420" alt=""/></div><br>
  <hr size="10" color="rgba(122,118,118,1.00)">
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    <div align="center"><img src="../../../网页/星级海景房旅游度假酒店商业网站模板/twts_79_royal/image/INTERLAB/Fig. 8 CFU Result1_meitu_9.jpg" width="732" height="492" alt=""/></div><br>
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  <p align="center" >Fig. 8 CFU Result </p></div>
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<div align="center"><h2 class="title_color">Discussion</h2></div>
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                <p>For Figure 3, the log graph isn’t a straight line but not 1:1 slope. In figure 6, highest fluorescence was obtained from device 4, closely followed by test device 1. Test device 2 and positive control group show a modest fluorescence intensity and device 5,6 show low fluorescence intensity, while test devices 3 barely have any fluorescence signal as well as the negative group.  
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<div align="center"><h2 class="title_color">Conclusion</h2></div>
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                 <p>It was certainly a technical challenge to Participate in the InterLab Study. Performing the prescribed protocols with adherence to all the InterLab guidelines yielded parts of expected results, and with the completed InterLab Google Forms, confirms our team participation in this InterLab Study.
                <div align="center"><h4 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2 . We discussed with the AHUT's model airplane team</h4></div>
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                  <p>Currently, our project is still at the laboratory stage, has not yet reached large-scale application. To better seek the scene of large-scale applications, Anhui University of Technology's model airplane team and we held a seminar and developed a program of social practice with the help of the instructor group. </p>
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/a/af/T--AHUT_China--_hangmo1.jpg" width="567" height="357" alt=""/></div><br>
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    <div align="center">Picture1 Group photo of Anhui University of Technology&#8217;s model airplane team</div>
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      We got in touch with a factory in Maanshan(Maanshan Steel Plant), and a six-rotor drone was provided by the model team, as shown in Picture 1<br><div align="center"><img src="https://static.igem.org/mediawiki/2018/4/4a/T--AHUT_China--_hangmo2.jpg" width="569" height="357" alt=""/></div>
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    <br><div align="center">Picture2 Six-rotor drone</div>
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    <p >Zhao Lei, who has studied embedded programming development in the team, has developed a carbon dioxide detection device that can be mounted on the unmanned aerial vehicle with different gas sensors (The gas sensor can detect the mass of carbon dioxide, sulfur dioxide and other gases in each liter.) and STM32 ARM microcontroller. As shown in Picture 3: </p>
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    <div align="center"><img src="https://static.igem.org/mediawiki/2018/7/79/T--AHUT_China--_hangmo3.jpg" width="567" height="357" alt=""/></div>
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  <div align="center">Picture 3 Gas detection device</div>
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<p >We used this drone to carry the detection device into the air and tested it near the end of the factory's exhaust system. It was found that the concentration of carbon dioxide near the smoke extraction device was extremely high, 5-20 times greater than the normal value. According to the document "National Greenhouse Gas Emissions Accounting and Reporting and Other 11 Greenhouse Gas Management National Standards Released", the proportion of various costs in the exhaust gas from such a plant's production process is: oxygen: carbon dioxide: sulfur dioxide: vulcanization Hydrogen: Carbon monoxide: Hydrogen chloride: Fluoride: Nitrogen oxides: Other = 14:10:3:3:3:2:3:8:54, <br><br>
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  We have measured the composition, content and mass percentage of each gas in the factory exhaust gas after averaging as follows: </p><hr size="10" color="rgba(68,64,64,1.00)">
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                <div align="center"><h4 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;3 . Cooperation with the Indian Institute of Technology </h4></div>
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                  <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;As the project continues to advance, HP's exchanges and cooperations between teams are more frequent and in-depth. After seeing the cooperation application issued by the iGEM team of the Indian Institute of Technology (IIT-Madras) at the iGEM official website, we contacted IIT-Madras to discuss cooperation actively. <br>
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                    We are pleased to collaborate with the iGEM team of the Indian Institute of Technology, Madras (IIT-Madras) to make a video on the science of synthetic biology in the mother tongue. The Indian Institute of Technology is also working with other teams in English, Tamil, Telugu, Kannada, Malayalam, Marathi, Bengali, Gujarati and Hindi. <br>
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                    &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;IIT-Madras sends an English script to our team. We need to match the provided slideshow video with the provided text and add the translated text to the subtitles, and finally make it into Chinese video. <br>
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                    &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We thought it was a simple matter, but there were some problems at the beginning. The problem is summarized as follows: <br>
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                    &amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;1. The English script does not exactly match the sound in the video, and some paragraphs are missing. <br>
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                    &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2. English and Chinese belong to different language families. They are slightly different in grammar, and the word order is reversed in translation. <br>
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                    &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;3. Some video pictures and sounds are out of sync and need to be adjusted. <br>
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                  &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 4. Our team members are not familiar enough with the Indian accent in English, and some words cannot be completely accurate. <br>
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                    &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;After consultation and repeated revisions by the iGEM team members of the Anhui University of Technology, four days later, the results were satisfactory and the video was sent to the iGEM team of the Indian Institute of Technology. We have established a good relationship with the iGEM team (IIT-Madras) of the Indian Institute of Technology, Madras. <br>
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                  With the successful completion of the HP cooperation, the Anhui University of Technology iGEM team (AHUT-CHINA) looks forward to more cooperation opportunities with the Indian Institute of Technology iGEM team (IIT-Madras). </p>
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<p align="center"><video controls="controls"><source src="https://static.igem.org/mediawiki/2018/2/21/T--AHUT_China--_indiaIITmp4.jpg" type="video/mp4"><p>
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                <div align="center"><h4 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;4 . Visit Hua Qi environmental companies (Zhou Yu) </h4></div>
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                  <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;On the afternoon of September 12, 2018, our team AHUT_China visited Maanshan Hua Qi Environmental Protection Technology Co., Ltd. (Hereinafter referred to as HuaQi Environmental Protection Company).</p>
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                  <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We know that environmental pollution is mainly air pollution, water pollution and soil pollution. Our project is to deal with greenhouse gas CO2 in air pollution. How to deal with greenhouse gas CO2 efficiently is particularly critical as the increasing greenhouse effect has a serious impact on the global environment. In order to gain a deeper understanding of the current environmental pollution in China, we visited HuaQi Environmental Protection Company. HuaQi Environmental Protection Company, is a company specializing in sewage treatment. After Ms. Zhang Qi introduced the company, we visited the intelligent water transportation management system and laboratory. </p>
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                  <div align="center"><img src="https://static.igem.org/mediawiki/2018/e/ec/T--AHUT_China--_Hua_Qi_environmental_companies1.jpg" width="466" height="365" alt=""/></div><br>
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                  <div align="center">Technical staff Liu Wei led us to visit the laboratory</div>
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  <p >&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Liu Shanshan, the company's technical staff, led us to visit the laboratory. During the visit, we learned that water pollution currently is harmful to the environment and people's health. So, water pollution treatment is very important. We introduced our project to computerize the protein molecules expressed by carbonic anhydrase gene mutations to obtain carbonic anhydrase (CA-2) with high thermal stability and import it into E. coli cells in order to achieve efficient collection of CO2. The project is currently in the theoretical stage. She is interested in our project. If the CO2 emission can be effectively controlled at the source, the problems such as the continued destruction of ozone layer, the melting of two-stage glaciers and the rapid extinction of a large number of species can be effectively solved. And she is more concerned about how this project can be applied in practice, and puts forward some suggestions for our project. At present, the CO2 gas capture technology mainly includes chemical absorption, pressure swing adsorption and membrane separation. She suggested first that transgenic E. coli be cultured in solution environment with chemical absorption to achieve CO2 collection, and secondly that CO2 be treated with biological rotating disc. </p>
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/0/06/T--AHUT_China--_Hua_Qi_environmental_companies2.jpg" width="466" height="365" alt=""/></div><br>
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                                  <div align="center">Group photo at Huaying Environmental Protection Company (second right: Liu Wei)</div>                
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                <div align="center"><h4 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;5 . <strong>iGEM(Linkoping_Sweden)</strong></h4></div>
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                  <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We saw the cooperation request issued by Linking University's iGEM team (Linkoping_Sweden). Linkoping_Sweden has a new idea of cooperation. They want other teams to use letters or emails to show interesting things happening inside and outside the team. So we participated in this cooperation. This is an interesting collaboration, as it may be boring during the experiment, but the stories that happen in the experiment make the experiment fun. And by organizing these stories, we learn more about our experiments. </p>
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                  <div align="center"><img src="https://static.igem.org/mediawiki/2018/0/03/T--AHUT_China--_Hua_Qi_environmental_companies3.jpg" width="794" height="442" alt=""/></div><br>
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                  <div align="center">Mail sent to Linköping University</div>
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  <div align="center"><img src="https://static.igem.org/mediawiki/2018/d/df/T--AHUT_China--_Hua_Qi_environmental_companies4.jpg" width="794" height="442" alt=""/></div>
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<div align="center">Reply email from Linköping University</div>
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                <div align="center"><h4 class="title_color">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;6. Biofilm Process Water Purification Laboratory                  </h4></div>
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                 <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;On September 8, 2018, our AHUT-China team visited the Biofilm Process Water Purification Laboratory of the College of Architecture and Engineering of Anhui University of Technology. The graduate student Xu Fan led us to visit the laboratory. The seniors introduced us to the principle of purifying sewage in the laboratory, which made us understand that relying on the microbial membrane fixed on the surface of the carrier to degrade organic matter has the characteristics of good effect and low technology and low cost.                 </p>
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                <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Then we introduced our project to the seniors. The seniors said that many problems caused by CO2 also brought us many problems. Therefore, he has great hope for our project. At the same time, we have learned that the biomembrance process can not only treat sewage, but also can be applied to the treatment of organic gases. The pollutants in the organic waste gas are first contacted with water and dissolved in water, that is, diffused from the gas film into the liquid film; The organic pollutant components in the liquid film are diffused to the biofilm by the difference in concentration, and they are captured and absorbed by the microorganisms therein; the organic pollutants entering the microorganisms are decomposed as energy and nutrients in the metabolism of the microorganisms themselves. This provides us with some ideas on the combination of theory and practice.                  </p>
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                <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Through a visit to the Biofilm Process Water Purification Laboratory in the afternoon, we learned about the biofilm treatment of sewage. At the same time, we also learned the necessity of our project. The impact of greenhouse gas CO2 cannot be underestimated. We also hope that our project can be applied to practice. The knowledge of biofilm processes in the treatment of organic waste gases has also given us some inspiration, and we may be able to use this knowledge when dealing with CO2. </p>
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              <div align="center"><img src="https://static.igem.org/mediawiki/2018/5/5b/T--AHUT_China--_Hua_Qi_environmental_companies5.jpg" width="536" height="398" alt=""/></div>
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<div align="center">Xu Fan, the head of the school, gave us a visit to the laboratory (second from the left: Xu Fan)</div><br>
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<div align="center"><img src="https://static.igem.org/mediawiki/2018/4/4a/T--AHUT_China--_Hua_Qi_environmental_companies6.jpg" width="536" height="398" alt=""/></div>
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                <div align="center">Taking a photo in the biofilm purification laboratory)</div><br>
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Revision as of 00:48, 5 October 2018

Royal Hotel Royal Hotel

     Introduction

We took part in the Fifth International InterLab Measurement Study which ains to achieve the purpose of comparative measurement. The goal of this study is to obtain large amounts of data from labs across the world,to develop absolute units for measurements GFP in a plate reader to eliminate variation between labs.


     Materials

Plate reader: Synergy H1 (Biotek)
Plate reader plates: Corning 3603 96-Well Microplates (black plates with clear flat bottom)
Cell culture shaker: ZWYR-200D

Devices:
Negative control :BBa_R0040
Positive control :BBa_I20270
Device 1: BBa_J364000
Device 2: BBa_J364001
Device 3: BBa_J364002
Device 4: BBa_J364007
Device 5: BBa_J364008
Device 6: BBa_J364009
Note: for Device 5, we have not transformed it into DH5⍺ competent cells successfully for many times, therefore, we thank IGEM team of Nanjing University for providing the Device 5.
Calibration material: Provided in the 2018 IGEM distribution kit
Microorganism: Escherichia coli DH5⍺ strains


     Methods

Following iGEM requirements, Team AHUT_China performed measurements according to these 2018 InterLab Protocols https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf


     Results

1. OD 600 reference point
Using OD 600 and H2O to generate the conversion factor for the transformation later. The average of OD600 is 0.063; the correction factor (OD600/ABS600) is 3.500



Fig. 1 LUDOX correct value

2.Particle standard curve
We obtained the two Particle Standard Curve (normal and log scale).



Fig. 2 Particle Standard Curve

2.Particle standard curve
We obtained the two Particle Standard Curve (normal and log scale).



Fig. 3 Particle Standard Curve (log scale)

3.Fluorescein standard curve
Dilution serious of fluorescein were prepared and measured in a 96 well plate. A standard curve is generated to correct the cell based readings to an equivalent fluorescein concentration.
We obtained the two Fluorescein Standard Curve (normal and log scale).



Fig. 4 Fluorescein Standard Curve

Fig. 5 Fluorescein Standard Curve (log scale)

4.Cell measurements

 



Fig. 6 Fluorescence Measurements Curve

Test devices 1 and 4 show high fluorescence intensity. Test devices 2 show a modest fluorescence intensity alone with positive control group, while devices3,5,6 barely show low fluorescence intensity alone with the negative control group.



Fig. 7 Raw OD600 Curve

5.We obtained the Colony Forming Units per 0.1 OD600 E. coli cultures




Fig. 8 CFU Result

 

Discussion

For Figure 3, the log graph isn’t a straight line but not 1:1 slope. In figure 6, highest fluorescence was obtained from device 4, closely followed by test device 1. Test device 2 and positive control group show a modest fluorescence intensity and device 5,6 show low fluorescence intensity, while test devices 3 barely have any fluorescence signal as well as the negative group.

Conclusion

It was certainly a technical challenge to Participate in the InterLab Study. Performing the prescribed protocols with adherence to all the InterLab guidelines yielded parts of expected results, and with the completed InterLab Google Forms, confirms our team participation in this InterLab Study.